Safety of beta‐nicotinamide mononucleotide (β‐NMN) pursuant the regulation (EU) 2015/2283 and the bioavailability of nicotinamide from this source in the context of Directive 2002/46/EC

Abstract

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on beta‐nicotinamide mononucleotide (β‐NMN) as a novel food (NF) pursuant to Regulation (EU) 2015/2283, including an evaluation of the safety of its use in food supplements as a source of niacin, and the bioavailability of nicotinamide from this source, in the context of Directive 2002/46/EC. The NF consists of chemically synthesised β‐NMN and is intended for use in food supplements as a source of niacin up to 300 mg/day. The target population is adults, excluding pregnant and lactating women. The identity, production process, composition and specifications of the NF do not raise safety concerns. A human intervention study comparing the relative bioavailability of β‐NMN and nicotinamide in equimolar amounts supported the use of a conversion factor (CF) of 1. On this basis, the proposed maximum intake corresponds to 109.7 mg/day of nicotinamide. This amount is approximately five times the population reference intake (PRI) of niacin for adults but remains below the upper level (UL) of 900 mg/day for nicotinamide. No concerns were identified regarding genotoxicity. In a 90‐day repeated dose oral toxicity study in rats, effect on reproductive organs were observed and the no observed adverse effect level (NOAEL) of 400 mg/kg bw per day identified, resulting in a margin of exposure (MoE) of 93. The available human study did not raise safety concerns. Based on the nature and the metabolic fate of the NF, the proposed use levels and the available human data, the calculated MoE was considered sufficient. The Panel concludes that the NF is safe under the proposed conditions of use for the adult population, excluding pregnant and lactating women and that the NF constitutes a bioavailable source of nicotinamide, a form of niacin.

1. INTRODUCTION

1.1. Background and Terms of Reference as provided by the requestor

The European Union legislation lists nutritional substances that may be used for nutritional purposes in certain categories of foods.

The relevant Union legislative measures are:

• Regulation (EU) 2015/2283 of the European Parliament and of the Council on novel foods. ¹

• Directive 2002/46/EC of the European Parliament and of the Council laying down requirements on food supplements. ²

On 2 August 2023, the company EffePharm (Shanghai) Co., Ltd. submitted an application to the European Commission in accordance with Article 10 of Regulation (EU) 2015/2283 to authorise the placing on the Union market of beta‐nicotinamide mononucleotide (β‐NMN) as a novel food.

The applicant requests to authorise the use of beta‐nicotinamide mononucleotide (β‐NMN) as a novel food in food supplements as defined by Directive 2002/46/EC, excluding food supplements for infants and young children.

The applicant has also requested data protection under Article 26 of Regulation (EU) 2015/2283.

In accordance with Article 29(l)(a) of Regulation (EC) No 178/2002, the European Commission asks the European Food Safety Authority to provide a scientific opinion:

• –by carrying out the assessment of beta‐nicotinamide mononucleotide (β‐NMN) as a novel food in accordance with Article 10(3) of Regulation (EU) 2015/2283, and
• –following the outcome of the novel food assessment, by evaluating the safety and bioavailability of beta‐nicotinamide mononucleotide (β‐NMN) when added for nutritional purposes to food supplements in the context of Directive 2002/46/EC, excluding food supplements for infants and young children, and
• –by assessing the extent to which beta‐nicotinamide mononucleotide (β‐NMN) is bioavailable as compared to native vitamin B3, as well as by deriving a conversion factor that allows converting absolute amounts of this substance into vitamin B3.

The Commission also asks EFSA to evaluate and inform the Commission as to whether and if so, to what extent, the requirements of Article 26(2)(c) of Regulation (EU) 2015/2283 are fulfilled in elaborating its opinion on beta‐nicotinamide mononucleotide (β‐NMN) as a novel food regarding the proprietary data for which the applicant is requesting data protection.

1.2. Information on existing evaluations and authorisations

In 2002, the Scientific Committee on Food (SCF) published an opinion on the Tolerable Upper Intake level (UL) for niacin [nicotinic acid (NA) and nicotinamide (NAM)] (SCF, 2022). An UL of 900 mg/day (12.5 mg/kg bw per day) was established for NAM for adults, excluding pregnant and lactating women in view of the lack of data for these population groups.

In 2014, the NDA Panel published an opinion on dietary reference values for niacin (EFSA NDA Panel, 2014).

2. DATA AND METHODOLOGIES

2.1. Data

The safety assessment of this NF is based on data supplied in the application and information submitted by the applicant following EFSA's requests for supplementary information.

Administrative and scientific requirements for NF applications referred to in Article 10 of Regulation (EU) 2015/2283 are listed in Commission Implementing Regulation (EU) 2017/2469. ³

A common and structured format on the presentation of NF applications is described in the EFSA guidance on the preparation and presentation of a NF application (EFSA NDA Panel, 2021). As indicated in this guidance, it is the duty of the applicant to provide all of the available (proprietary, confidential and published) scientific data (including both data in favour and not in favour), that are pertinent to the safety of the NF.

The applicant has submitted a confidential and a non‐confidential version of a dossier, following the ‘EFSA guidelines on the preparation and presentation of a NF application’ (EFSA NDA Panel, 2021) and the ‘Administrative guidance for the preparation of applications on novel foods pursuant to Article 10 of Regulation (EU) 2015/2283’ (EFSA, 2021).

In accordance with Article 38 of Regulation (EC) No 178/2002 ⁴ and taking into account the protection of confidential information and of personal data in accordance with Articles 39 to 39e of the same Regulation, and of the Decision of EFSA's Executive Director laying down practical arrangements concerning transparency and confidentiality, ⁵ the non‐confidential version of the dossier has been published on Open.EFSA. ⁶

According to Article 32c(2) of Regulation (EC) No 178/2002 and to the Decision of EFSA's Executive Director laying down the practical arrangements on pre‐submission phase and public consultations, EFSA carried out a public consultation on the non‐confidential version of the technical dossier from 4th February to 25th February 2025. The comments received have been carefully evaluated and considered during the current assessment (Annex A).

This NF application includes a request for protection of proprietary data in accordance with Article 26 of Regulation (EU) 2015/2283. The studies requested by the applicant to be protected are listed in Appendix B.

2.2. Methodologies

The assessment follows the methodology set out in the EFSA guidance on NF applications (EFSA NDA Panel, 2021) and the principles described in the relevant existing guidance documents from the EFSA Scientific Committee. The legal provisions for the assessment are laid down in Article 11 of Regulation (EU) 2015/2283 and in Article 7 of Commission Implementing Regulation (EU) 2017/2469.

This assessment concerns only the risks that might be associated with consumption of the NF under the proposed conditions of use and is not an assessment of the efficacy of the NF with regard to any claimed benefit.

The evaluation of bioavailability of the nutrient niacin from the source beta‐nicotinamide mononucleotide was conducted in line with the principles contained in the ‘Guidance on safety evaluation of sources of nutrients and bioavailability of nutrient from the sources’ (EFSA ANS Panel, 2021).

3. ASSESSMENT

3.1. Introduction

The NF which is subject of the application is a chemically synthetised compound and according to the applicant, as defined by Article 3 of the NF Regulation (EU) 2015/2283, falls under the category (2)(a)(i) food with a new or intentionally modified molecular structure, where that structure was not used as, or in, a food within the Union before 15 May 1997.

The NF is the chemical compound beta‐nicotinamide mononucleotide (β‐NMN) produced by chemical synthesis in a multi‐step process and it consists of a crystalline powder (purity ≥ 99%). The NF is proposed to be used in food supplements as a source of niacin at daily doses up to 300 mg. The target population is adults, excluding pregnant and lactating women.

3.2. Identity of the NF

The NF, β‐NMN, is a chemical compound with a purity of ≥ 99%. The chemical identity and the chemical structure of the NF, as provided by the applicant, are reported in Table 1 and in Figure 1, respectively.

TABLE 1.

Chemical identity of the NF.

Chemical substance
Chemical (IUPAC) name	3‐Carbamoyl‐1‐[5‐O‐(hydroxyphosphinato)‐β‐D‐ribofuranosyl]pyridinium or [(2R,3S,4R,5R)‐5‐(3‐carbamoylpyridin‐1‐ium‐1‐yl)‐3,4‐dihydroxyoxolan‐2‐yl]methyl dihydrogen phosphate
Common name	Nicotinamide mononucleotide
Synonyms, trade names, abbreviations	Beta‐nicotinamide mononucleotide; beta‐NMN; β‐NMN; nicotinamide ribonucleoside 5′‐phosphate; β‐nicotinamide riboside monophosphate; 3‐(aminocarbonyl)‐1‐(5‐O‐phosphonato‐beta‐D‐ribofuranosyl)pyridinium
CAS number	1094‐61‐7
Molecular formula	C11H15N2O8P
Molecular weight	334.221 g/mol

FIGURE 1.

Chemical structure of the NF, beta‐nicotinamide mononucleotide.

The applicant provided a characterisation of the NF by multiple identification methods. Specifically, the identity of the NF was characterised through HPLC with a diode array detector, mass and infrared spectrometric analysis and proton (¹H) and carbon‐13 (¹³C) nuclear magnetic resonance (NMR). The beta configuration was demonstrated by direct comparison with an authentic standard, with values reported in the literature as well as confirmed using ¹H‐¹H correlation spectroscopy. Upon EFSA's request the applicant also provided optical rotation measurements, comparing the results directly with an authentic reference standard to support the identity and purity of the NF.

The water solubility of the NF was investigated according to OECD TG 105, using the flask method with ultrafiltration as requested by the relevant Guidance (EFSA Scientific Committee, 2021). The test result showed that the NF is highly soluble in water (i.e. 167 g/L) and it does not raise concerns regarding consumer exposure to small particles.

3.3. Production process

According to the information provided, the NF is produced in line with Good Manufacturing Practice (GMP) and Hazard Analysis Critical Control Points (HACCP) principles.

The NF is produced by chemical synthesis in a multi‐step process under controlled conditions. In the first step of the reaction process, nicotinamide riboside chloride (NR‐Cl), trimethyl phosphate (TMP) and phosphorus oxychloride (POCl₃) react together to yield nicotinamide mononucleotide‐chloride (NMN‐Cl). Thereafter, NMN‐Cl is subjected to a hydrolysis reaction to yield β‐NMN, which is then purified by ion‐exchange chromatography, filtered and crystalised to obtain the final ingredient.

The Panel considers that the production process is sufficiently described.

3.4. Compositional data

The NF consists of a white to off‐white crystalline powder with a purity of ≥ 99%.

In order to confirm that the manufacturing process is reproducible and adequate to produce on a commercial scale a product with certain characteristics, the applicant provided analytical information for five independently produced batches of the NF (Table 2).

TABLE 2.

Batch‐to‐batch analysis of the NF.

Parameter (unit)	Batch 1	Batch 2	Batch 3	Batch 4	Batch 5	Method of analysis
Assay (% on a dry basis)	99.43	99.42	99.40	99.21	99.37	HPLC (internal method)
Purity (%)	99.87	99.72	99.67	99.64	99.67	HPLC (internal method)
Water (%)	0.29	0.33	0.26	0.15	0.22	Karl Fischer
	0.24	0.24	0.21	0.21	0.20	GB 5009.3‐2016
Specific optical rotation (about 5% aqueous solution, w/v) (°)	−50.676	−50.018	−50.349	−50.318	−49.711	General Rule 0621 of the Chinese Pharmacopoeia (2020 edition) Part Four: Determination of Optical Rotation
Metals and other elements
Lead (mg/kg)	0.024	0.038	0.024	0.022	< 0.02 a	ICP‐MS (GB 5009.268‐2016)
Cadmium (mg/kg)	< 0.002 a	< 0.002 a	< 0.002 a	< 0.002 a	< 0.002 a
Total arsenic (mg/kg)	0.009	0.013	0.004	0.016	0.019
Mercury (mg/kg)	< 0.001 a	< 0.001 a	< 0.001 a	< 0.001 a	< 0.001 a
Microbiological parameters
Total viable plate count (CFU/g)	< 10 a	< 10 a	< 10 a	< 10 a	< 10 a	USP43‐NF38 (2020) Ch. 61
Yeasts and moulds (CFU/g)	< 10 a	< 10 a	< 10 a	< 10 a	< 10 a
Salmonella in 25 g	Not detected	Not detected	Not detected	Not detected	Not detected	AOAC 2003.09 BAX
Escherichia coli in 10 g	Not detected	Not detected	Not detected	Not detected	Not detected	ISO 7251:2005+Amd1:2023 detection method
Coagulase positive staphylococci in 10 g	Not detected	Not detected	Not detected	Not detected	Not detected	ISO 6888‐3:2003 (E) detection method
Coliforms (negative/1 g)	< 0.3 a	< 0.3 a	< 0.3 a	< 0.3 a	< 0.3 a	GB 4789.30‐2016
Residual solvent and reagents
Ethanol (mg/kg)	69	67	85	77	65	Chinese Pharmacopoeia 2020 Part IV general rule (0861)
Ethyl acetate (mg/kg)	< 50 b	< 50 b	< 50 b	< 50 b	< 50 b
Trimethyl phosphate (mg/kg)	< 0.5 a	< 0.5 a	< 0.5 a	< 0.5 a	< 0.5 a
Phosphorus oxychloride (%)	< 0.002 a	< 0.002 a	< 0.002 a	< 0.002 a	< 0.002 a	HG/T3606‐2009 (Phosphoryl chloride for industrial use)

Abbreviations: AOAC, Association of Official Analytical Collaboration; CFU, colony forming units; GB, Guobiao (Chinese National Standard); HPLC, high‐performance liquid chromatography; ICP–MS, inductively coupled plasma–mass spectrometry; ISO, International Organization for Standardization; USP, United States Pharmacopeia; w/v, weight per volume.

^a LOD.

^b LOQ.

According to the data submitted by the applicant, the NF assay and purity was determined using an in‐house method based on HPLC, yielding values above 99% and with a water content up to 0.33%.

Metals and other elements such as lead, cadmium, total arsenic and mercury were analysed through inductively coupled plasma–mass spectrometry (ICP–MS) with results below the respective LODs for cadmium and mercury (0.002 and 0.001 mg/kg, respectively), while being up to 0.038 and 0.019 mg/kg for lead and total arsenic, respectively. All results were in compliance with Commission Regulation (EC) 2023/915 when applicable.

Microbiological parameters were also conducted according to the United States Pharmacopeia, ISO and AOAC analytical methods. The results showed that total aerobic counts and total yeast and mould counts were both below 10 CFU/g, while Escherichia coli and coagulase positive Staphylococci were not detected in 10 g and Salmonella was not detected in 25 g.

Residual solvents and reagents were analysed according to the procedures of the Chinese Pharmacopeia. All results were below their respective LODs, except for ethanol (Table 2).

Information was provided on the accreditation of the laboratories that conducted the analyses presented in the application.

The Panel considers that the information provided on the composition is sufficient for characterising the NF.

3.4.1. Stability

The NF is intended to be stored at room temperature in sealed containers. The intended shelf life is 24 months. The applicant performed stability tests with five independently produced batches of the NF. The tests were carried out at accelerated conditions at 40°C and at 75% relative humidity (RH) for a period of 6 months. The batches were analysed for identity, appearance, purity, water content, microbiological parameters, metals and other elements, and residual solvents. Three additional batches were also analysed under real‐time conditions at 25°C and 60% RH at 0 and 24 months, analysed for identity, purity, water content and microbiological parameters. In both accelerated and real‐time studies, the tested parameters remained unchanged.

The Panel considers that the data provided sufficient information with respect to the stability of the NF.

3.5. Specifications

The specifications of the NF are indicated in Table 3.

TABLE 3.

Specifications of the NF.

Description: β‐NMN is an odourless, white to off‐white crystalline powder with a chemical purity of more than 99% and is highly soluble in water
Parameter	Specification
Identity and composition
Assay (%, on a dry basis)	99.00–102.00
Purity (%)	≥ 99.0
Water (%)	≤ 1.0
Specific optical rotation (about 5% aqueous solution, w/v) (°)	−49° to −51°
Residual solvents
Ethyl acetate (mg/kg)	≤ 50
Ethanol (mg/kg)	≤ 100
Trimethyl phosphate (mg/kg)	≤ 0.5
Phosphorus oxychloride (%)	≤ 0.002
Metals and other elements 7
Total arsenic (mg/kg)	≤ 0.05
Microbiological parameters
TAMC	≤ 1000 (CFU/g)
TYMC	≤ 100 (CFU/g)
Salmonella in 25 g	Not detected
Escherichia coli in 10 g	Not detected
Coagulase positive staphylococci in 10 g	Not detected
Coliforms (MPN/g)	< 0.92

Abbreviations: CFU, colony forming units; MPN, most probable number; TAMC, total aerobic microbial count; TYMC, total yeast and mould count; w/v, weight per volume.

The applicant did not originally propose a specification limit for POCl₃. However, based on the results of the batch‐to‐batch analyses of the novel food and in order to ensure its safety, the Panel considers it appropriate that a maximum limit for POCl₃ is included in the specifications and proposes that this limit is set at ≤ 0.002% POCl₃.

In addition, while the applicant proposed the determination of optical rotation in an aqueous solution at 1%, the Panel considers it appropriate to include the parameter specification at 5% to ensure consistency with the batch‐to‐batch analytical data and for which a certificate of analysis was provided.

The Panel considers that the information provided on the specifications of the NF is sufficient and does not raise safety concerns.

3.6. History of use of the NF and/or of its source

3.6.1. History of use of the source

The starting material for the NF is NR‐Cl. In 2019, the NDA Panel established the safety of NR‐Cl as a novel food pursuant to Regulation (EU) 2015/2283 for use in food supplements for the adult population, and the bioavailability of nicotinamide from this source, in the context of Directive 2002/46/EC (EFSA NDA Panel, 2019).

In 2021, the NDA Panel evaluated an extension of use of NR‐Cl to include meal replacement products and nutritional drink mixes (up to 300 mg/day for the general population), as well as foods for special medical purposes (FSMP) and total diet replacement for weight control (TDRWC) (up to 500 mg/day in adults). The safety of NR‐Cl for use in FSMP and TDRWC at the proposed doses was confirmed, whereas the safety could not be established for meal replacement products and nutritional drink mixes under the proposed conditions of use (EFSA NDA Panel, 2021).

3.6.2. History of use of the NF

Outside the European Union, the NF has been subject to different regulatory classifications. In Canada, it is regulated as a Natural Health Product under the Natural Health Products Regulations. ⁸

In the US, β‐NMN was notified to the U.S. Food and Drug Administration (FDA) as a new dietary ingredient intended for use in food supplements in 2022. However, the agency considered it excluded from the dietary supplement definition on the basis that β‐NMN had been previously investigated as a drug product. ⁹ In 2025, the FDA reconsidered its position and concluded that β‐NMN can be considered as a dietary supplement. ¹⁰

According to the applicant, the NF has been sold in form of a food supplement since 2019 in Australia and Hong Kong, and since 2020 in South Korea, Japan, New Zealand, Taiwan and Canada.

3.7. Proposed uses and use levels and anticipated intake

3.7.1. Target population

The target population proposed by the applicant is adults, excluding pregnant and lactating women.

3.7.2. Proposed uses and use levels

The applicant intends to market the NF for use in food supplements, at a maximum dose of 300 mg per day, which corresponds to an equimolar amount of 109.7 mg nicotinamide per day.

3.8. Absorption, distribution, metabolism and excretion (ADME)

The hydrolysis of the NF as a function of pH was investigated by the applicant by dissolving 1 g of β‐NMN in 100 mL of a buffer solution with a pH of either 4, 7 or 9 at 50°C, which were incubated for 5 days (Unpublished study, 2023a). The NF was hydrolysed by less than 10% in all three samples, suggesting stability in that pH range. The Panel notes that mean gastric pH in an empty (fasting) stomach is lower (around 1.5–2) and that these pH levels were not tested.

The applicant acknowledges that the pharmacokinetics of β‐NMN are not completely known. Based on data on the uptake into/translocation across intestinal cell models under simulated GI digestion conditions using in vitro systems, the applicant proposes that absorption of β‐NMN could occur directly via a transporter located on the apical side of the intestinal epithelium as intact β‐NMN, or indirectly by dephosphorylation into nicotinamide riboside (NR) via the ecto‐5′‐nucleotidase CD73 (Grozio et al., 2019; Smith and Brenner, 2019; Kim et al., 2023).

Once absorbed and present in the systemic circulation, and owing to the lack of a transporter, the NF could be metabolised extracellularly to NR, which is then taken up by cells and converted back to β‐NMN by the enzyme nicotinamide riboside kinase via a phosphorylation reaction (Poddar et al., 2019; Yoshino et al., 2018), before being metabolised into nicotinamide adenine dinucleotide (NAD) (Poddar et al., 2019). The NF is thus proposed to have a niacin function as a precursor of NAD and nicotinamide adenine dinucleotide phosphate (NADP), and a similar metabolic fate.

The major pathway of catabolism of dietary niacin (nicotinic acid and nicotinamide) is by methylation in the liver to N‐methyl‐nicotinamide and subsequent oxidation to N‐methyl‐2‐pyridone‐carboxamide (2‐Pyr) and N‐methyl‐4‐pyridone‐carboxamide (4‐Pyr). In humans, the two major excretion products in urine are N‐methyl‐nicotinamide and 2‐Pyr, which under normal conditions represent about 20%–35% and 45%–60% of niacin metabolites, respectively. The amount of niacin metabolites excreted in urine depends on the niacin and tryptophan intake (EFSA NDA Panel, 2014).

3.8.1. Bioavailability of the NF

Bioavailability of the NF after oral administration has been assessed in mice using isotope‐labelled β‐NMN (Mills et al., 2016; Kim et al., 2022). These studies suggest that the NF is quickly absorbed intact after oral consumption and is effectively used for NAD synthesis in cells. Another study in rats, in which nicotinamide or β‐NMN were intraperitoneally injected and the concentration of NAD in blood and of NAD metabolites in urine were measured over 12h, suggests that metabolism and excretion of β‐NMN may be delayed vs. that of nicotinamide (Kawamura et al., 2016).

Human intervention studies, including one conducted by the applicant (Huang et al., 2022), have investigated the effect of the NF at doses up to 900 mg/day given for up to 12 weeks on plasma levels of β‐NMN and/or NAD and its metabolites as compared to a placebo, with variable results (Igarashi et al., 2022; Irie et al., 2020; Katayoshi et al., 2023; Okabe et al., 2022; Yi et al., 2023). Blood concentrations of NAD and its metabolites significantly increased in all the studies following consumption of β‐NMN as compared to placebo. Blood concentrations of β‐NMN significantly increased or were unchanged, depending on the study (Igarashi et al., 2022; Katayoshi et al., 2023; Okabe et al., 2022). The Panel notes that, although these studies provide some evidence that the NF is absorbed by humans upon oral consumption and may be used as a precursor of NAD, they do not inform about the relative bioavailability of the NF proposed as a new nutrient source as compared to an appropriate reference source, e.g. nicotinamide (EFSA NDA Panel, 2024).

3.8.2. Relative bioavailability of the NF and derivation of a conversion factor for labelling purposes

The applicant provided an in vitro bidirectional permeability study using Caco‐2 cells (Unpublished study, 2024a), a study in male Sprague–Dawley rats (Unpublished study, 2024b) and a human intervention study comparing β‐NMN and nicotinamide (Unpublished study, 2025).

From the in vitro study, the applicant concluded that the permeability rate across Caco‐2 cells is higher for β‐NMN than for nicotinamide, and that neither substance is a potential substrate for efflux transporters (Unpublished study, 2024a). In the rat study, a total of 12 fasted male Sprague–Dawley rats (3 per group) were administered 1496 μmol/kg bw of either β‐NMN or nicotinamide (single dose), either orally (gavage) or intravenously (i.v.). Blood samples were collected via jugular vein cannulation at the following time points: pre‐dose, 5 (i.v. only), 15 and 30 min, and 1, 2, 4, 6, 8, 12 and 24 h post dose. Concentrations of β‐NMN and nicotinamide in plasma were determined by liquid chromatography–tandem mass spectrometry (LC–MS/MS). ■■■■■ The Panel considers that, although these two studies further support bioavailability of the NF upon oral consumption, they are of limited value to derive a conversion factor for β‐NMN as compared to nicotinamide.■■■■■

The human intervention study (Unpublished study, 2025) was initially conducted as an open‐label, cross‐over randomised controlled trial (RCT) in which 10 adult Asian volunteers [8 males and 2 females; 6 vegetarian and 4 non‐vegetarian; age range: 28–40 years; Body Mass Index (BMI) range: 18.3–28.6 kg/m²] consumed 100, 300 and 600 mg β‐NMN, and 109.7 mg nicotinamide (the equimolar amount for 300 mg β‐NMN dose) in a random order, with a washout of 7 days between each two doses. Upon EFSA's request to include equimolar amounts of nicotinamide for the 100 and 600 mg β‐NMN doses (36.5 and 219.2 mg, respectively), the applicant simply extended the study providing these two additional nicotinamide doses in a random order, with a 7‐day washout in between. The Panel notes that the consumption of the substances and doses tested was only partially randomised, and that neither participants nor laboratory personnel were blinded to the interventions.

During the study, participants were provided with a standard low‐niacin diet to consume at home. The study products were taken on test days after an overnight fast. Blood was drawn at baseline and 1, 2, 4, 8, 12 and 24‐h post‐administration for the measurement of plasma and erythrocyte concentrations of NAD, and plasma concentrations of β‐NMN, nicotinamide, N‐methyl‐nicotinamide, 2‐Pyr and 4‐Pyr. Concentrations of nicotinamide, N‐methyl‐nicotinamide, 2‐Pyr, 4‐Pyr and creatinine were measured in timed 24‐h urine collections obtained from participants on each test day. The results were provided for each test product and tested dose by time point in blood and urine. The C _max and AUC for each parameter (for different time periods and over 24 h) were also calculated. Upon EFSA's request, the applicant also provided the results for each metabolite and for all metabolites combined for the entire 24‐h urine collections, both as concentrations and in absolute values using total urine volumes.

The Panel considers that, even if the C _max and the time at which the C _max is achieved for the different metabolites provides useful information about the different time profiles of β‐NMN and nicotinamide in increasing plasma and urine concentrations of nicotinamide, NAD and its metabolites, the 24‐h AUC and total 24‐h urinary excretion of all metabolites, individually and combined, best reflect the relative bioavailability of β‐NMN vs. nicotinamide. The results for these parameters, and the respective β‐NMN/nicotinamide ratio of means thereof that could be used to derive a conversion factor for β‐NMN, were provided by the applicant.

■■■■■

Taking into account that niacin metabolites are excreted in urine even at low‐niacin intakes, that plasma niacin metabolites are less sensitive to changes in niacin intakes than urinary metabolites, and that for intakes > 11 mg/day urinary excretion of niacin metabolites reflects saturation of body stores (EFSA NDA Panel, 2014), the Panel considers that a conversion factor of 1 can be derived for supplemental β‐NMN vs. nicotinamide across the dose range of 100–600 mg/day when consumed as a single dose in water, based on the data provided by the applicant.

3.9. Nutritional information

The NF consists of β‐NMN (≥ 99%) and is proposed to be used as a source of niacin in food supplements intended for adults (≥ 18 years of age), excluding pregnant and lactating women.

Niacin is a generic term for nicotinic acid and nicotinamide, which are water‐soluble organic compounds that belong to the group of B vitamins. In vivo, nicotinic acid is converted to nicotinamide, and thus both compounds are identical in their vitamin function. Niacin can be obtained from food or synthetised in the liver from the indispensable amino acid tryptophan. Approximately 60 mg of tryptophan yields 1 mg of niacin, defined as 1 mg niacin equivalent (NE). The function of niacin is as the precursor of NAD and NADP, which are involved in oxidation/reduction reactions and associated with both catabolic and anabolic processes (EFSA NDA Panel, 2014).

Since niacin requirements are dependent on energy intake, Dietary Reference Values for niacin have been established on that basis (EFSA NDA Panel, 2014). A Population Reference Intake (PRI) of 1.6 mg NE/MJ (about 6.6 mg NE/1000 kcal) has been derived for adults, corresponding to daily intakes between 10.9 and 22.4 mg/day, considering average requirements (AR) for energy derived for adult men and non‐pregnant, non‐lactating women at different physical activity levels (PAL) (EFSA NDA Panel, 2013). Separate tolerable upper intake levels (UL) for free nicotinic acid and nicotinamide have also been established (SCF, 2022). The Panel considers that only the UL for nicotinamide of 900 mg/day established for adults is relevant for this safety assessment.

The Panel considers that, taking into account the composition of the NF and the proposed conditions of use, consumption of the NF is not nutritionally disadvantageous.

3.10. Toxicological information

The applicant provided several toxicological studies conducted with the NF. These studies are listed in Table 4.

TABLE 4.

List of toxicological studies with the NF.

References	Type of study	Test system	Dose
Unpublished study (2021a)	Bacterial reverse mutation test (GLP, OECD TG 471)	S. Typhimurium TA 1535, TA97a, TA98, TA100 and TA102	Up to 5000 μg/plate (absence and presence of S9 mix)
Unpublished study (2023b)	In vitro mammalian cell micronucleus test (GLP, OECD TG 487)	CHO‐K1 cell line	Up to 2000 μg/mL (absence and presence of S9 mix)
Unpublished study (2021b)	In vitro mammalian chromosomal aberration test (GLP, OECD TG 473)	CHO cell line	Up to 2000 mg/kg μg/mL (absence and presence of S9 mix)
Unpublished study (2021c)	In vivo mammalian Bone Marrow Chromosomal Aberration Test (GLP, OECD TG 475)	Swiss albino mice	500, 1000 and 2000 mg/kg bw
Unpublished study (2021d)	90‐day repeated dose oral toxicity study with a 28‐day recovery period (GLP, OECD TG 408)	Wistar rats	0, 200, 400 and 800 mg/kg bw per day

Abbreviations: bw, body weight; CHO, Chinese hamster ovary; GLP, Good Laboratory Practice; OECD, Organization for Economic Co‐operation and Development; TG, test guideline.

The applicant states that the specifications of β‐NMN used in the toxicological studies has minor differences from those proposed for the novel food, notably with respect to purity (≥ 99.0% for the NF vs. ≥ 98.0% for the test material), water content (≤ 1.0% vs. ≤ 5.0%) and arsenic levels (≤ 0.5 mg/kg vs. ≤ 1 mg/kg). The Panel considers the β‐NMN used in the toxicological studies to represent a worst‐case scenario, as the proposed NF specifications reflect higher purity and lower impurity limits.

3.10.1. Genotoxicity

A bacterial reverse mutation assay (Ames test) was performed to evaluate the potential of β‐NMN (purity 98.9%) to induce gene mutations in S. Typhimurium strains TA97a, TA98, TA100, TA102, TA1535. The study was carried out in accordance with OECD test guideline (TG) 471 (1997, corrected in 2020) and was claimed to be compliant with Good Laboratory Practice (GLP). Two independent experiments were performed applying the plate incorporation and pre‐incubation methods in the presence and absence of metabolic activation (S9‐mix). The test item, dissolved in distilled water, was tested using five concentrations ranging from 50 to 5000 μg/plate. No precipitation or cytotoxicity, as indicated by a reduction in the background lawn, was observed in any tester strain and experimental condition. The historical negative control data were based on DMSO (not distilled water) but were considered sufficient to confirm the validity of the assay and the functionality of the S9 metabolic activation system. No significant changes in the number of revertant colonies were induced by the test item compared to the vehicle control under any experimental condition.

The Panel considers that β‐NMN did not induce gene mutations in bacteria under the experimental conditions employed in this study.

An in vitro micronucleus test was carried out to evaluate the potential of the NF (purity 99.7%) to induce chromosomal damage in CHO (Chinese Hamster Ovary) cell line. The study was carried out in accordance with OECD TG 487 (2023) and was claimed to be compliant with GLP. The test item, dissolved in distilled water, was used at 500, 1000, 2000 μg/mL, applying a short treatment in the presence and absence of S9‐mix (4 + 17 h of recovery) and a continuous treatment in the absence of S9‐mix (21 + 0 h of recovery). No increased cytotoxicity was observed under any experimental condition. No statistically significant increase in the frequency of micronucleated cells was observed at any concentration and sampling time. Positive and negative control values were within historical laboratory data, confirming assay validity and the functional integrity of the S9 metabolic activation system. The Panel concludes that the test item did not induce structural and numerical chromosomal damage in mammalian cells under the experimental conditions employed.

The β‐NMN (purity 98.9%) was also evaluated for its potential to induce chromosomal damage in the Chinese Hamster Ovary (CHO‐K1) cell line in a study carried out in accordance with OECD TG 473 (2016a) and claimed to be compliant with GLP. The test item, dissolved in distilled water, was used at 500, 1000, 2000 μg/mL, applying a short treatment in the presence and absence of S9 (3 + 17 h of recovery) and a continuous treatment in the absence of S9 (20 + 0 h of recovery). No cytotoxicity was induced by the test item under any experimental condition. No significant changes in the frequency of structural chromosome aberrations, polyploid and endoreduplicated cells were observed at any concentration and sampling time in comparison to the vehicle controls. The Panel concludes that the test item did not induce structural chromosome aberrations in mammalian cells under the experimental conditions employed in the study.

The genotoxicity potential of the NF was further investigated in an in vivo mammalian bone marrow chromosomal aberration test performed in Swiss albino mice according to OECD TG 475 (OECD, 2016b) to evaluate the potential of the NF (purity 98.9%) to induce structural chromosome aberrations. A dose‐range finding study, performed in male and female animals, showed no clinical sign of toxicity or mortality in any of the treated animals and no differences between males and females. On this basis, in the main study, the test item (dissolved in distilled water) was administered as a single treatment by gavage at 500, 1000, 2000 mg/kg bw only to female mice. Animals were sacrificed 24 h after dosing. No toxicity was observed in the bone marrow in the treated groups relative to the negative control mice, as measured by the mitotic index. The frequency of chromosome aberrations was comparable between treated and negative control groups. Therefore, the Panel concluded that the test item did not induce structural chromosome aberrations under the experimental conditions employed in this study.

Taking into account the test results provided and considering the nature, source and production process of the NF, the Panel considers that there are no concerns regarding genotoxicity.

3.10.2. Subchronic toxicity

The applicant provided a 90‐day repeated dose oral toxicity study in Wistar rats with a 28‐day recovery period conducted according to OECD TG 408.

The NF was administered by gavage at doses of 0 (distilled water), 200, 400 and 800 mg/kg bw per day (corresponding to NAM equivalent to 73, 146 and 292 mg/kg bw per day) to both male and female Wistar rats (10/sex per group) for 90 days, followed by a 28‐day recovery (5/sex per group) for groups 0 and 800 mg/kg bw per day.

There were no clinical signs and no significant effects in the functional observation battery, and no mortalities during the study. Statistically significant decreases in body weight were observed at the mid and high dose on days 85 and 90. However, the changes were less than 10% and therefore not considered adverse. No changes in food consumption were observed.

A statistically significant increase was observed in the following haematological parameters. In males, only the mid dose induced an increase in neutrophils (+48%), monocytes (+69%) and a decrease in lymphocytes (−17%) was observed. An increase in basophils (+53%) was observed at the high dose. All changes observed in males were not dose‐dependent.

In females, an increase in neutrophil counts was observed at the mid and high dose level (+49 and +69%, respectively) accompanied by a decrease in lymphocytes (−16 and −19%, respectively). Red blood cell counts (+7%) and haemoglobin (+9%) were increased at the high dose. Haematocrit slightly increased at mid (6%) and high (7%) dose. At the mid dose, mean corpuscular volume (MCV) increased slightly (+3%). The changes in the red blood cell parameters were below 10% and were considered as not toxicologically relevant. Platelet counts (PLT) decreased at mid (−14%) and high (−11%) dose. All these changes were dose‐dependent except for PLT. The Panel notes that in the recovery groups, no significant differences were observed between treated and control animals. In addition, all the significant changes recorded at the end of the 90‐day treatment period were comparable to those of the recovery control group.

Based on the above considerations, the Panel concludes that the observed effects are unlikely to be associated with the test material and are not considered to be adverse.

In males, a significant decrease in total serum bilirubin was observed only at the mid dose (−31%). A reduction in creatinine (−11%) at the high dose was noted, and cholinesterase activity (+57%) and albumin levels (+6%) were increased at the low dose only, without a consistent pattern across dose groups. Serum lactate dehydrogenase (LDH) and serum aspartate aminotransferase (AST) were decreased at the high dose (−32% and −16%, respectively). A decrease in total protein was observed at the mid dose (−14%). Reductions in chloride at mid (−3%) and high (−4%) doses and increase of sodium at the high (2%) doses were minimal and considered not of biological relevance.

In females, a decrease in total cholesterol was observed at the high dose (−19%) as well as a decrease in cholinesterase activity (−23%) at the low dose. An increase in LDH (+44%) and AST (+23%) was observed at the mid dose. Minor increases in sodium at mid (+5%) and high doses (+3%), as well as decreases in chloride across all dose groups (−2%, −3% and −4% respectively), remained within physiological variability and are therefore considered not treatment‐related.

Overall, the Panel considers that most of the observed alterations in clinical chemistry parameters in both sexes were generally small in magnitude and considered as not treatment‐related.

In males, an increase in absolute brain weight was observed at the high dose (+11%). Statistically significant increases were also reported for the seminal vesicles plus prostate, both in absolute (+40%) and relative (+42%) weight at the high dose. Similarly, epididymides weights were elevated at the high dose, with increases of 19% (absolute) and 17% (relative to body weight). These changes were outside of the historical control range. The Panel notes that in the male recovery group a significant decrease in heart weight relative to body weight was observed at the highest dose after 28‐day recovery (−18%).

In females, thymus weight relative to body weight was increased at the high dose (+32%), while ovarian weight relative to body weight was decreased (−22%). An increase in uterine weight relative to body weight (+4%) was also noted at the high dose. Additionally, a decrease in absolute thyroid and parathyroid weights was observed at mid and high doses (−34 and −22% respectively). The Panel notes that in the female recovery group a significant decrease in kidney absolute and relative body weight was observed at the highest dose after 28‐day recovery (−17% and −15%, respectively).

No histopathological findings were reported.

The Panel considers the changes in the reproductive organs in the high‐dose group as adverse and establishes a no observed adverse effect level (NOAEL) of 400 mg/kg bw per day, i.e. the mid‐dose group of the study.

3.10.3. Human data

The applicant submitted a multicentre, randomised, double‐blind, parallel, placebo‐controlled study performed in accordance with Good Clinical Practice (Huang, 2022; Unpublished study, 2021e). Males and females from 40 to 65 years of age with a BMI between 18.5 and 35 kg/m² were administered either one 150 mg β‐NMN (NF) capsule twice daily (total daily dose of 300 mg) or a placebo capsule (containing starch powder) twice daily for 60 days. Out of 70 participants screened, 66 met the inclusion/exclusion criteria and were randomised. The statistical analysis was conducted in the 62 subjects completing the intervention (n = 31 per group).

The primary objective was to investigate the effect of β‐NMN supplementation on several efficacy endpoints. The secondary objective was to assess the safety and tolerability of the formulation. Several laboratory parameters, including biochemistry, haematology and urinalysis, were assessed at screening and end of study.

There were no changes observed in the safety endpoints measured, and no serious adverse events were reported. Dyslipidaemia was reported in one subject from the placebo group and one from the treated group.

Other published randomised placebo‐controlled, human intervention studies using highly pure β‐NMN produced with different production processes are summarised in Appendix A. Six trials were conducted in adult populations comprising healthy subjects (Fukamizu et al., 2022; Igarashi et al., 2022; Katayoshi et al., 2023; Okabe et al., 2022; Yi et al., 2023) or adults with type 2 diabetes (Akasaka et al., 2023). Across the studies, both males and females were included, with participants aged ≥ 20 years. Sample sizes ranged from 7 to 20 subjects in the β‐NMN arms. The interventions lasted from 4 to 24 weeks and investigated daily doses between 250 and 1250 mg β‐NMN. Safety‐related endpoints commonly comprised clinical chemistry parameters, haematology, urinalysis, vital signs and monitoring of adverse events. No treatment‐related serious adverse events were reported and no clinically relevant differences between treatment and placebo groups were observed on the parameters investigated. Overall, the Panel notes that these studies did not identify adverse effects associated with the consumption of highly pure β‐NMN under the investigated conditions.

The Panel notes that no adverse effects of β‐NMN were reported in the above‐mentioned human intervention studies, which tested doses up to 900 mg/day for 60 days and up to 250 mg/day for 24 weeks. Study populations comprised men and women, mostly middle aged (40–65 years) or older adults (> 65 years). The Panel considers that these human data are relevant for the safety assessment, that the study populations are representative of the intended adult population and that they capture part of the variability within that population.

The Panel concludes that the available human data support the safety of the NF.

3.11. Allergenicity

The Panel considers that, owing to the nature of the NF and the production process, which does not involve proteins, it is unlikely that the NF will trigger allergic reactions in the target population under the proposed conditions of use.

4. DISCUSSION

The NF subject to the application is beta‐nicotinamide mononucleotide (β‐NMN; purity ≥ 99.0%) produced by chemical synthesis. The NF is proposed to be used in food supplements as a source of niacin at daily doses up to 300 mg. The target population is adults, excluding pregnant and lactating women.

The Panel considers that the information provided on the identity, production process, composition and specifications of the NF is sufficient and does not raise safety concerns.

β‐NMN is bioavailable in humans, is a precursor of nicotinamide adenine dinucleotide and is metabolised and excreted through similar pathways as nicotinamide. The information provided by the applicant was sufficient to assess the relative bioavailability of the NF as a source of niacin in humans, and to derive a CF of 1 as compared to nicotinamide under the proposed conditions of use.

The maximum use levels of 300 mg/day of the NF as food supplement corresponds to an equimolar amount of 109.7 mg nicotinamide per day.

A NOAEL of 400 mg/kg bw per day for the NF was identified for the 90‐day repeated dose oral toxicity study in rats, corresponding to a margin of exposure (MoE) of 93 for adults.

A PRI of 1.6 mg NE/MJ for niacin has been derived for adults (EFSA NDA Panel, 2014), corresponding to daily intakes between 10.9 and 22.4 mg/day. The UL for nicotinamide for adults is 900 mg/day for adults (SCF, 2022). The maximum use level proposed for the NF corresponds to a nicotinamide intake which is about five times the PRI of niacin for adults, but below the UL for nicotinamide of 900 mg/day.

The human intervention studies conducted with the NF or highly pure β‐NMN in adult men and women, at doses up to 900 mg/day for 60 days and up to 250 mg/day for 24 weeks, did not identify adverse effects and support the safety of the NF under the proposed conditions of use.

Based on the nature and the metabolic fate of the NF, the proposed use levels that are below the UL for nicotinamide, the available human data and the MoE of 93 for adults derived from the 90‐day toxicological study which the Panel considers to be sufficient in this case, the Panel concludes that the NF does not raise safety concerns under the proposed conditions of use.

5. CONCLUSIONS

The Panel considers that the NF, β‐NMN, is safe under the proposed conditions of use. The Panel concludes that the NF is a source from which niacin is bioavailable.

5.1. Protection of Proprietary data in accordance with Article 26 of Regulation (EU) 2015/2283

The Panel could not have reached the conclusion on the safety of the NF under the proposed conditions of use without the data claimed as proprietary by the applicant as listed in Appendix B.

Abbreviations

2‐Pyr

N‐methyl‐2‐pyridone‐carboxamide

4‐Pyr

N‐methyl‐4‐pyridone‐carboxamide

A/G

albumin/globulin ratio

ADME

absorption, distribution, metabolism and excretion

AE

adverse event

ANS

Panel on Food Additives and Nutrient Sources added to Food

AOAC

Association of Official Analytical Collaboration

AR

average requirements

AST

aspartate aminotransferase

AUC

area under the curve

BMI

body mass index

bw

body weight

CAS

Chemical Abstracts Service

CF

conversion factor

CFU

colony forming unit

CHO

Chinese Hamster Ovary

C _max

maximum concentration

DMSO

dimethyl sulfoxide

FDA

U.S. Food and Drug Administration

FSMP

Foods For Special Medical Purposes

GB

Guobiao (Chinese National Standard)

GI

gastrointestinal

GLP

Good Laboratory Practice

GMP

Good Manufacturing Practice

HACCP

hazard analysis critical control points

HDLC

high‐density lipoprotein cholesterol

HPLC

high‐performance liquid chromatography

i.v.

intravenously

ICP–MS

inductively coupled plasma–mass spectrometry

ISO

International Organization for Standardization

IUPAC

International Union of Pure and Applied Chemistry

LC–MS/MS

liquid chromatography–tandem mass spectrometry

LDH

serum lactate dehydrogenase

LOD

limit of detection

LOQ

limit of quantification

MCHC

mean corpuscular haemoglobin concentration

MCV

mean corpuscular volume

MoE

margin of exposure

MPN

most probable number

NA

nicotinic acid

NAD

nicotinamide adenine dinucleotide

NADP

nicotinamide adenine dinucleotide phosphate

NAM

nicotinamide

NDA

EFSA Panel on Nutrition, Novel Foods and Food Allergens

NDI

new dietary ingredient

NE

niacin equivalent

NF

novel food

NMN‐Cl

nicotinamide mononucleotide‐chloride

NMR

nuclear magnetic resonance

NOAEL

no observed adverse effect level

NR

nicotinamide riboside

NR‐Cl

nicotinamide riboside chloride

OECD TG

Organisation for Economic Co‐Operation and Development Test Guidelines

PAL

physical activity levels

PLT

platelet counts

POCl₃

phosphorus oxychloride

PRI

population reference intake

RCT

randomised controlled trial

RH

relative humidity

S9

metabolic activation system

SCF

Scientific Committee on Food

TAMC

total aerobic microbial count

TDRWC

total diet replacement for weight control

TMP

trimethyl phosphate

TYMC

total yeast and mould count

UL

tolerable upper intake levels

USP

United States Pharmacopeia

w/v

weight per volume

β‐NMN

beta‐nicotinamide mononucleotide

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2023‐00552

COPYRIGHT FOR NON‐EFSA CONTENT

EFSA may include images or other content for which it does not hold copyright. In such cases, EFSA indicates the copyright holder and users should seek permission to reproduce the content from the original source.

PANEL MEMBERS

Dominique Turck, Torsten Bohn, Montaña Cámara, Jacqueline Castenmiller, Stefaan De Henauw, Karen Ildico Hirsch‐Ernst, Ángeles Jos, Alexandre Maciuk, Inge Mangelsdorf, Breige McNulty, Androniki Naska, Kristina Pentieva, Alfonso Siani and Frank Thies.

LEGAL NOTICE

Relevant information or parts of this scientific output have been blackened in accordance with the confidentiality requests formulated by the applicant pending a decision thereon by EFSA. The full output has been shared with the European Commission, EU Member States (if applicable) and the applicant. The blackening may be subject to review once the decision on the confidentiality requests is adopted by EFSA and in case it rejects some of the confidentiality requests.

Supporting information

Annex A: Public consultation on the draft scientific opinion on the safety of beta‐nicotinamide mononucleotide (β‐NMN) pursuant the regulation (EU) 2015/2283 and the bioavailability of nicotinamide from this source in the context of Directive 2002/46/EC

APPENDIX A. Summary characteristics of the human intervention studies conducted using highly pure β‐NMN (oral route)

References	Study design	Study population	Duration of study	Dose	Safety‐related parameters investigated
Fukamizu et al. (2022)	Randomised, double‐blinded placebo‐controlled, parallel‐group study	31 healthy subjects (20–65 years) of which 15 placebo (8 women and 7 men; BMI 22.1 ± 3.3) 16 treated (7 men and 9 women; BMI 22.9 ± 2.7)	4 weeks	1250 mg	Haematology (RBC, Hb, Ht, WBC, PLT), clinical biochemistry [liver (AST, ALT, ALP, TBil); kidney (Cr, BUN, UA); metabolism (Glu, TC, TG); electrolytes (Na, K, Cl, Ca); other enzymes (LDH, CK), BW, BMI; vital signs (BP, HR, BT), urinalysis (Prot, Glu, OB)] and adverse events were assessed at baseline, Week 2, Week 4 and 2 weeks post‐treatment. No clinically relevant differences were identified between‐ or within‐groups on the parameters tested. No AEs related to the treatment.
Yi et al. (2023)	Multicentre, randomised, parallel, double‐blinded, placebo‐controlled, dose‐dependent study	80 heathy men and women (49.3 ± 6.8 years; BMI 27 ± 5) (20 placebo and 20 for each treatment dose)	60 days	300, 600 or 900 mg/day	General and systemic examinations, vital signs, clinical assessments (BP, HR, BT, ECG, chest Xray), haematology (Hb, Ht‐PCV, MCHC, MCV, RBC, WBC, PT, aPTT), clinical chemistry [liver (AST, ALT, ALP, TBil, Alb, Prot); kidney (Cr, urea, BUN, GFR, UA); metabolism (Glu, TC, LDL, HDL, TG, HOMA‐IR); electrolytes (Na, K, Cl, Ca)] and urinalysis (pH, SG, PRO, GLU, KET, LEU, NIT, Hb/RBC, UBG, BIL) were assessed at baseline and Day 60. No drop‐out due to treatment. No clinically relevant differences were identified between treatments and placebo groups on the parameters tested. No AEs related to the treatment.
Igarashi et al. (2022)	Placebo‐controlled, randomised, double‐blind, parallel‐group study	42 healthy older men (> 65 years of age; BMI 24 ± 2) (21 placebo and 21 treated)	12 weeks	250 mg/day	Haematology (WBC, RBC, Hb, Ht, PLT, MCH, MCV, MCHC); clinical chemistry [liver (AST, ALT, γ‐GTP); kidney (Cr, BUN, UA); electrolytes (Na, K); metabolism (FBG, HbA1c, Insulin, C‐peptide, HOMAIR, HOMAβ, Glu AUC, Insulin AUC, C‐peptide; AUC; TG, TC, LDL‐cholesterol, HDLC); others: CK, hsCRP, Adiponectin, IL6, Alb, TP] were assessed at Week 0 and 12. No drop‐out due to treatment. No clinically relevant differences were identified between treatment and placebo groups on the parameters tested. No AEs related to the treatment.
Okabe et al. (2022)	Placebo‐controlled, randomised, double‐blind, parallel‐group study	30 healthy men and women (20–65 years; BMI 21 ± 3) (15 placebo and 15 treated)	12 weeks	250 mg/day	Haematology (WBC, RBC, Hb, Ht, PLT, MCV, MCH, MCHC); clinical chemistry [liver (AST, ALT, γGTP, ALP); kidney (Cr, BUN, UA); electrolytes and minerals (Na, K, Cl, Ca, IP, Mg, Fe); metabolism (FPG, HbA1c, IRI, HOMA‐IR, HOMAβ; TG, TC, LDL‐cholesterol, HDLC, FFA, Lp(a); AcAc, 3‐HBA, TKB); other (LDH, TP, Alb)]; urinalysis (Glu, Prot, UBG, Ket, pH, SG, OB, Bil); vital signs and anthropometrics (BW, BMI, SBP, DBP, PR) and adverse events, were assessed repeatedly from baseline to Week 12, with followup at Week 16. One drop‐out in placebo group. No clinically relevant differences were identified between treatment and placebo groups on the parameters tested. No AEs related to the treatment.
Katayoshi et al. (2023)	Randomised, double‐blind, placebo‐controlled, parallel‐group	36 healthy men and women (40–59 years of age; BMI 22 ± 4) (18 placebo and 18 treated)	12 weeks	250 mg/day	Haematology (WBC, RBC, Hb, Ht); clinical chemistry [liver (AST, ALT, γGTP); metabolism (blood glucose, TG, LDL‐cholesterol, HDLC); hormones (testosterone, progesterone, estradiol, DHEAS, cortisol)]; vital signs and anthropometrics (BW, BMI, SBP, DBP) were assessed at baseline and after 12 weeks of intervention. No clinically relevant differences were identified between treatment and placebo groups on the parameters tested. AEs not reported.
Akasaka et al. (2023)	Placebo‐controlled, double‐blinded	15 men with type 2 diabetes (≥ 65 years of age; BMI NR) (7 placebo and 6 treated)	24 weeks	250 mg/day	Clinical chemistry (GLU, HbA1c, CRP); vital signs and anthropometrics (BW, SBP, DBP) and adverse events were assessed at baseline, 12 weeks and 24 weeks. Two drop‐outs due to AEs, one in placebo and one in treatment group. No differences were reported between treatment and placebo groups on the parameters tested (data not shown). No AEs related to the treatment.

Note: Mean ± SD.

Abbreviations: 3‐HBA, 3‐hydroxybutyric acid; γGTP, gammaglutamyl transpeptidase; A/G, albumin/globulin ratio; AcAC, acetoacetic acid; AEs, adverse events; ALP, alkaline phosphatase; ALT, alanine aminotransferase; aPTT, activated partial thromboplastin time; AST, aspartate aminotransferase; BIL, bilirubin; BMI, body mass index; BP, blood pressure; BT, body temperature; BUN, blood urea nitrogen; BW, body weight; Ca, calcium; CK, creatine kinase; Cl, chloride; Cr, creatinine; CRP, C‐reactive protein; DBP, diastolic blood pressure; DHEAS, dehydroepiandrosterone sulfate; ECG, electrocardiogram; FFA, free fatty acid; FPG, fasting plasma glucose; GLU, glucose; Hb, haemoglobin; Hb/RBC, haemoglobin/erythrocytes (urine); HbA1c, haemoglobin A1c; HDLC, High‐density lipoprotein cholesterol; HOMA‐IR, homeostasis model assessment‐insulin resistance; HR, heart rate; hsCRP, high‐sensitivity C‐reactive protein; Ht, haematocrit; Ht‐PCV, haematocrit‐packed cell volume; IL6, Interleukin‐6; K, potassium; KET, ketones; LDH, lactate dehydrogenase; LDL, low‐density lipoprotein; LEU, leukocytes; Lp(a), lipoprotein (a); MCH, mean corpuscular haemoglobin; MCHC, mean corpuscular haemoglobin concentration; MCHC, mean corpuscular haemoglobin concentration; MCV, mean corpuscular volume; Na, sodium; NIT, nitrite; NR, not reported; OB, urinary occult blood; PLT, platelet count; PR, pulse rate; PRO, protein; Prot, urinary protein; RBC, red blood cell count; SBP, systolic blood pressure; SD, standard deviation; SG, specific gravity; TBil, total bilirubin; TC, total cholesterol; TG, triglycerides; TKB, total ketone bodies; TP, total protein; UA, uric acid; UBG, urobilinogen; WBC, white blood cell count.

APPENDIX B. List of elements of the dossier for which data protection was requested

NF section	Elements of the application dossier for which a request for data protection was filed by the applicant	File considered as pertinent for the NDA Panel to reach the conclusion on the safety (yes/no)
Identity	Solubility study report (Annex E)	Yes
Compositional data	Certificate of analysis (Annexes D, Annexes H)	Yes
	Method validation (Annex G)	No
History of use	FDA correspondence (Annexes A)	No
ADME	Hydrolysis as a function of pH (Annex P)	No
	Bidirectional permeability and efflux ration in Caco‐2 cells (Annex S)	No
	Pharmacokinetic comparison study (Annex T)	No
	Human pharmacokinetic study (Annex U)	Yes
Toxicological information	Bacterial reverse mutation test (Annex I, Genotoxicity)	Yes
	In vitro micronucleus assay (Annex R, Genotoxicity)	Yes
	In vitro chromosomal aberration assay (Annex J, Genotoxicity)	Yes
	In vivo chromosomal aberration assay (Annex K, Genotoxicity)	Yes
	Acute oral toxicity (Annex L, Acute toxicity)	No
	Repeated dose 90 day study (Annex M, Subchronic toxicity)	Yes
	Clinical trial (Annex N, Human Data)	Yes

EFSA NDA Panel (EFSA Panel on Nutrition, Novel Foods and Food Allergens) , Turck, D. , Bohn, T. , Cámara, M. , Castenmiller, J. , De Henauw, S. , Jos, Á. , Maciuk, A. , Mangelsdorf, I. , McNulty, B. , Naska, A. , Pentieva, K. , Siani, A. , Thies, F. , Aguilera‐Gómez, M. , Cubadda, F. , Frenzel, T. , Gundert‐Remy, U. , Marcon, F. , … Hirsch‐Ernst, K. I. (2026). Safety of beta‐nicotinamide mononucleotide (β‐NMN) pursuant the regulation (EU) 2015/2283 and the bioavailability of nicotinamide from this source in the context of Directive 2002/46/EC . EFSA Journal, 24(5), e10007. 10.2903/j.efsa.2026.10007